Glycocholicacid, glycodeoxycholicacid, glycochenodeoxycholicacid, taurocholicacid, taurodeoxycholicacid and taurochenodeoxycholicacid are identified for the first time in intact human bile using several 1D and 2D [1H-1H and 1H-13C] experiments at 400, 700 and 800 MHz and a method is proposed for their accurate analyses. 1H and 13C chemical shifts for ten standard bile acids were determined under physiological conditions and used for unambiguous identification of bile acids in the complex mixture of human bile. Simple one step method for accurate analyses of individual bile acids in bile presented herein may have implications in the study of bile acid metabolism associated with biliary diseases.